Fig. 5

Efficient lysosome labeling is obtained using SiR-Tet while MVB labeling requires TAMRA-Tet. a–c Cells expressing GCE-tag-Lamp1 labeled with SiR-Tet (a) or Lamp1-mVenus (b). Left panels: no treatment. Right panels: treatment with the lysosome inhibitor chloroquine (3 h, 120 μM). Zoomed in images of a subset of the cells (correspond to squares in upper panels) are shown in bottom panels. Arrows indicate filled versus hollow lysosome labeling. c Left to right: line intensity profiles measured for the red dashed line in a, the green dashed line in b, and a summary of SNR values measured in cells expressing the indicated plasmids (n = 50). d COS7 cells expressing the MVB marker CD63 conjugated to the GCE-tag were fixed and stained with anti-HA and anti-CD63 antibodies. e, f MVB labeling in cells expressing GCE-tag-CD63 labeled with TAMRA-Tet (e) or CD63-mCherry (f). g SNR values measured in cells expressing the indicated plasmids. n = 50. Results presented in each panel were obtained in at least 3 independent experiments. Scale-bars, 10 μm; zoomed-in images, 2 μm